STOCK Cdk6^tm2.1Phin Cdk4^tm1Bbd/Cnbc

Status	Available to order
EMMA ID	EM:06842
Citation information	RRID:IMSR_EM:06842 Research Resource Identifiers (RRID) are persistent unique ID numbers assigned to help researchers cite key resources (e.g. antibodies, model organisms and software projects) in the biomedical literature to improve transparency and reproducibility in research. See https://www.rrids.org/ for more information.
International strain name	STOCK Cdk6^tm2.1Phin Cdk4^tm1Bbd/Cnbc
Alternative name	Cdk 4R24C;Cdk 6R31C or Cdk4 tm1Bbd;Cdk6 R31C
Strain type	Targeted Mutant Strains : Knock-in
Allele/Transgene symbol	Cdk4^tm1Bbd, Cdk6^tm2.1Phin
Gene/Transgene symbol	Cdk4, Cdk6

Information from provider

Provider	Mariano Barbacid
Provider affiliation	Molecular Oncology, Centro Nacional de Investigaciones Oncologicas
Additional owner	Co-ownership by Dr. Mariano Barbacid and Dr. Marcos Malumbres, Centro Nacional de Investigaciones Oncologicas, Madrid, Spain
Genetic information	A loxP-flanked neo resistance cassette was inserted between Ex1 and 2 of Cdk4. In addition, the AAGCCCGA sequence in Ex2 (nt81-88) was replaced by AAGCTTGT, altering codon 24 from one corresponding to Arg to Cys. A third silent substitution at position nt85 was used to create a diagnostic restriction enzyme site to monitor the presence of the mutation. Germline cre recombinase-mediated excision of the neo cassette results in the expression of the mutated Cdk4^R24C protein. This aminoacid substitution prevents the binding of Cdk4 to INK4 inhibitors. A loxP-flanked PGK-neo cassette was integrated in the first intron of Cdk6. In addition, we replaced the GCCCGC (Ala30-Arg31) sequence in Ex1 by GCATGC. The point mutation in codon 31 (CGC to TGC) results in the replacement of a wild-type Arg residue by a Cys (R31C). This mutation prevents the binding of Cdk6 to INK4 inhibitors. Germ-line cre-mediated excision of the neo cassette results in the expression of the mutated Cdk6^R31C protein.
Phenotypic information	Expression of the Cdk4^R24C mutant protein leads to spontaneous tumor development, with near 100% penetrance and multiple tumors per animal, both in heterozygosis and homozygosis. Most tumors have an endocrine origin: insulinomas, pituitary adenomas and Leydig cell tumors in testis. Cdk6^R31C mice display defective maturation of lymphocytes and increased susceptibility to lymphoma development. Double mutant mice exhibit a combination of both phenotypes.
Breeding history	The two mutations were created by independent gene targeting. Chimeras carrying the Cdk4^R24C allele were crossed with CD1 females for germ line transmission analysis. Cdk4^R24C heterozygous animals were crossed with Tg(CMV-cre)1Nagy for deleting the neomycin resistance cassette. Chimeras carrying the Cdk6^R31C allele were crossed with CD1 females for germ line transmission analysis. Heterozygous offspring were crossed with Tg(CMV-cre)1Nagy for deleting the neomycin resistance cassette. Cdk4^R24C and Cdk6^R31C heterozygous animals were inter-crossed. Line has been maintained by mating homozygous animals for both alleles. Genetic background involves 129S1/Sv,129X1/SvJ and CD1.
References	Mammalian cells cycle without the D-type cyclin-dependent kinases Cdk4 and Cdk6.;Malumbres Marcos, Sotillo Rocío, Santamaría David, Galán Javier, Cerezo Ana, Ortega Sagrario, Dubus Pierre, Barbacid Mariano, ;2004;Cell;118;493-504; 15315761 Loss of Cdk4 expression causes insulin-deficient diabetes and Cdk4 activation results in beta-islet cell hyperplasia.;Rane S G, Dubus P, Mettus R V, Galbreath E J, Boden G, Reddy E P, Barbacid M, ;1999;Nature genetics;22;44-52; 10319860 Wide spectrum of tumors in knock-in mice carrying a Cdk4 protein insensitive to INK4 inhibitors.;Sotillo R, Dubus P, Martín J, de la Cueva E, Ortega S, Malumbres M, Barbacid M, ;2001;The EMBO journal;20;6637-47; 11726500 Genetic rescue of Cdk4 null mice restores pancreatic beta-cell proliferation but not homeostatic cell number.;Martín Javier, Hunt Sarah L, Dubus Pierre, Sotillo Rocío, Néhmé-Pélluard Fanny, Magnuson Mark A, Parlow Albert F, Malumbres Marcos, Ortega Sagrario, Barbacid Mariano, ;2003;Oncogene;22;5261-9; 12917627 Postnatal Schwann cell proliferation but not myelination is strictly and uniquely dependent on cyclin-dependent kinase 4 (cdk4).;Atanasoski Suzana, Boentert Matthias, De Ventura Lukas, Pohl Hartmut, Baranek Constanze, Beier Konstantin, Young Peter, Barbacid Mariano, Suter Ueli, ;2008;Molecular and cellular neurosciences;37;519-27; 18191580
Homozygous fertile	yes
Homozygous viable	yes
Homozygous matings required	no
Immunocompromised	no

Information from EMMA

Archiving centre	CNB-CSIC, Centro Nacional de Biotecnologia, Madrid, Spain
Animals used for archiving	heterozygous mixed males

Disease and phenotype information

Orphanet associated rare diseases, based on orthologous gene matching

Autosomal recessive primary microcephaly / Orphanet_2512

IMPC phenotypes (gene matching)

hypoplasia / IMPC
abnormal embryo size / IMPC
preweaning lethality, incomplete penetrance / IMPC
developmental dysplasia / IMPC
abnormal eye morphology / IMPC
decreased body length / IMPC
increased mean corpuscular volume / IMPC
edema / IMPC
female infertility / IMPC
microphthalmia / IMPC
male infertility / IMPC
increased mean corpuscular volume / IMPC
small superior vagus ganglion / IMPC
abnormal cranium morphology / IMPC
increased mean corpuscular hemoglobin / IMPC

MGI phenotypes (allele matching)

abnormal female reproductive system morphology / MGI
small ovary / MGI
abnormal male reproductive system morphology / MGI
decreased body weight / MGI
decreased body size / MGI
abnormal locomotor behavior / MGI
hyperactivity / MGI
polydipsia / MGI
impaired limb coordination / MGI
hyperglycemia / MGI
increased urine glucose level / MGI
polyuria / MGI
reduced male fertility / MGI
female infertility / MGI
postnatal lethality / MGI
abnormal seminiferous tubule morphology / MGI
abnormal corpus luteum morphology / MGI
oligozoospermia / MGI
decreased circulating insulin level / MGI
abnormal Sertoli cell morphology / MGI
decreased circulating follicle stimulating hormone level / MGI
ketoaciduria / MGI
abnormal cell cycle / MGI
abnormal male reproductive system physiology / MGI
abnormal female reproductive system physiology / MGI
decreased testis weight / MGI
decreased circulating progesterone level / MGI
abnormal pancreatic islet morphology / MGI
Leydig cell hypoplasia / MGI
delayed estrous cycle / MGI
small pancreatic islets / MGI
small thymus / MGI
decreased thymocyte number / MGI
abnormal T cell differentiation / MGI
decreased hematopoietic stem cell number / MGI
increased double-negative T cell number / MGI
decreased double-positive T cell number / MGI
increased CD4-positive, alpha beta T cell number / MGI
increased CD8-positive, alpha-beta T cell number / MGI
decreased DN1 thymic pro-T cell number / MGI
decreased DN2 thymocyte number / MGI
increased DN3 thymocyte number / MGI
decreased DN4 thymocyte number / MGI
abnormal hematopoietic stem cell physiology / MGI

MGI phenotypes (gene matching)

abnormal female reproductive system morphology / MGI
small ovary / MGI
abnormal male reproductive system morphology / MGI
abnormal testis morphology / MGI
small testis / MGI
Leydig cell hyperplasia / MGI
small seminiferous tubules / MGI
increased body weight / MGI
decreased body weight / MGI
decreased body size / MGI
abnormal locomotor behavior / MGI
hyperactivity / MGI
polydipsia / MGI
impaired limb coordination / MGI
hyperglycemia / MGI
increased urine glucose level / MGI
polyuria / MGI
reduced male fertility / MGI
male infertility / MGI
female infertility / MGI
abnormal glucose homeostasis / MGI
postnatal lethality / MGI
abnormal fertility/fecundity / MGI
no abnormal phenotype detected / MGI
abnormal seminiferous tubule morphology / MGI
abnormal corpus luteum morphology / MGI
oligozoospermia / MGI
decreased circulating insulin level / MGI
abnormal Sertoli cell morphology / MGI
absent Leydig cells / MGI
decreased circulating follicle stimulating hormone level / MGI
ketoaciduria / MGI
abnormal cell cycle / MGI
impaired luteinization / MGI
increased pancreatic beta cell number / MGI
abnormal male reproductive system physiology / MGI
abnormal female reproductive system physiology / MGI
decreased testis weight / MGI
azoospermia / MGI
decreased circulating progesterone level / MGI
abnormal pancreatic islet morphology / MGI
homeostasis/metabolism phenotype / MGI
Leydig cell hypoplasia / MGI
decreased lactotroph cell number / MGI
adenohypophysis hypoplasia / MGI
delayed estrous cycle / MGI
small pancreatic islets / MGI
postnatal lethality, incomplete penetrance / MGI
decreased leukocyte cell number / MGI
abnormal spleen morphology / MGI
small spleen / MGI
spleen hypoplasia / MGI
abnormal thymus morphology / MGI
small thymus / MGI
enlarged thymus / MGI
increased thymocyte number / MGI
decreased thymocyte number / MGI
decreased body weight / MGI
thymus hypoplasia / MGI
reduced female fertility / MGI
abnormal definitive hematopoiesis / MGI
abnormal T cell differentiation / MGI
abnormal spleen red pulp morphology / MGI
abnormal hematopoietic system morphology/development / MGI
abnormal lymphopoiesis / MGI
abnormal erythrocyte morphology / MGI
increased mean corpuscular volume / MGI
decreased mature ovarian follicle number / MGI
decreased erythrocyte cell number / MGI
thymus atrophy / MGI
abnormal thymocyte activation / MGI
increased hematopoietic stem cell number / MGI
decreased hematopoietic stem cell number / MGI
decreased double-negative T cell number / MGI
increased double-negative T cell number / MGI
decreased double-positive T cell number / MGI
decreased T cell proliferation / MGI
immune system phenotype / MGI
increased mean corpuscular hemoglobin concentration / MGI
increased T cell apoptosis / MGI
increased CD4-positive, alpha beta T cell number / MGI
increased CD8-positive, alpha-beta T cell number / MGI
decreased CD8-positive, alpha-beta T cell number / MGI
increased single-positive T cell number / MGI
decreased megakaryocyte cell number / MGI
decreased thymocyte apoptosis / MGI
increased DN1 thymic pro-T cell number / MGI
decreased DN1 thymic pro-T cell number / MGI
decreased DN2 thymocyte number / MGI
increased DN3 thymocyte number / MGI
decreased DN4 thymocyte number / MGI
abnormal hematopoietic stem cell physiology / MGI
thymus cortex atrophy / MGI

Literature references

Mammalian cells cycle without the D-type cyclin-dependent kinases Cdk4 and Cdk6.;Malumbres Marcos, Sotillo Rocío, Santamaría David, Galán Javier, Cerezo Ana, Ortega Sagrario, Dubus Pierre, Barbacid Mariano, ;2004;Cell;118;493-504; 15315761
Loss of Cdk4 expression causes insulin-deficient diabetes and Cdk4 activation results in beta-islet cell hyperplasia.;Rane S G, Dubus P, Mettus R V, Galbreath E J, Boden G, Reddy E P, Barbacid M, ;1999;Nature genetics;22;44-52; 10319860
Wide spectrum of tumors in knock-in mice carrying a Cdk4 protein insensitive to INK4 inhibitors.;Sotillo R, Dubus P, Martín J, de la Cueva E, Ortega S, Malumbres M, Barbacid M, ;2001;The EMBO journal;20;6637-47; 11726500
Genetic rescue of Cdk4 null mice restores pancreatic beta-cell proliferation but not homeostatic cell number.;Martín Javier, Hunt Sarah L, Dubus Pierre, Sotillo Rocío, Néhmé-Pélluard Fanny, Magnuson Mark A, Parlow Albert F, Malumbres Marcos, Ortega Sagrario, Barbacid Mariano, ;2003;Oncogene;22;5261-9; 12917627
Postnatal Schwann cell proliferation but not myelination is strictly and uniquely dependent on cyclin-dependent kinase 4 (cdk4).;Atanasoski Suzana, Boentert Matthias, De Ventura Lukas, Pohl Hartmut, Baranek Constanze, Beier Konstantin, Young Peter, Barbacid Mariano, Suter Ueli, ;2008;Molecular and cellular neurosciences;37;519-27; 18191580

Information on how we integrate external resources can be found here

Order

Availabilities

Requesting frozen sperm or embryos is generally advisable wherever possible, in order to minimise the shipment of live mice.

Frozen sperm. Delivered in 4 weeks (after paperwork in place). €1740^*
Rederivation of mice from frozen stock, delivery time available upon request . €3880^*

Due to the dynamic nature of our processes strain availability may change at short notice. The local repository manager will advise you in these circumstances.

^* In addition users have to cover all the shipping costs (including the cost for returning dry-shippers, where applicable).

More details on pricing and delivery times

Practical information

Genotyping protocol

Example health report
(Current health report will be provided later)

Material Transfer Agreement (MTA)
Distribution of this strain is subject to a provider MTA. Both signing of the MTA and submission of the online EMMA Mutant Request Form are required before material can be shipped.

EMMA conditions
Legally binding conditions for the transfer

Right strain for your research?

The information provided on this page is, to the best of EMMA’s knowledge, based on data supplied by the original provider. End users are responsible for reviewing these details and for validating the strain and its suitability for their experimental use.
Not found what you were looking for? Search here for other strains available from EMMA.

STOCK Cdk6tm2.1Phin Cdk4tm1Bbd/Cnbc